[Evaluation of the performance of InnowaveDX MTB/RIF for the detection of Mycobacterium tuberculosis complex and rifampicin resistance].

Objective: To evaluate the performance of Mycobacterium tuberculosis and rifampicin resistance mutation detection kit (InnowaveDX MTB/RIF, referred to as "InnowaveDX") in diagnosing tuberculosis and rifampicin resistance using sputum samples. Methods: From June 19, 2020 to May 16, 2022, patients with suspected tuberculosis were prospectively and consecutively enrolled in Hunan Provincial Tuberculosis Prevention and Control Institute, Henan Provincial Hospital of Infectious Diseases and Wuhan Jinyintan Hospital. A total of 1 328 patients with suspected tuberculosis were finally included. According to the inclusion and exclusion criteria, 1 035 pulmonary tuberculosis patients (357 were confirmed tuberculosis cases and 678 were clinically diagnosed tuberculosis cases) and 180 non-tuberculosis patients were finally included. Sputum samples were collected from all patients for routine sputum smear acid-fastness tests, mycobacterial culture and drug susceptibility testing. Moreover, the diagnostic value of Xpert®MTB/RIF (referred to as "Xpert") and InnowaveDXin detecting tuberculosis and rifampicin resistance was evaluated. Clinical diagnosis and culture results of Mycobacterium tuberculosis were used as reference standards to assess tuberculosis diagnosis, and phenotypic drug sensitivity and Xpert were used as reference standards to assess rifampicin resistance. The sensitivity, specificity, positive predictive value and negative predictive value of the two methods for tuberculosis diagnosis and rifampicin resistance were analyzed. The consistency of the two techniques was analyzed usingkappa test. Results: Taking clinical diagnosis as the reference standard, the detection sensitivity of InnowaveDX [58.0% (600/1 035)] was higher than that of Xpert [51.7% (535/1 035)] in 1035 patients with pulmonary tuberculosis, and the difference was statistically significant (P<0.001). In 270 pulmonary tuberculosis patients with culture-positive pulmonary tuberculosis identified as M.tuberculosis-complex, the positive rates of InnowaveDX and Xpert were both high [99.6%(269/270)and 98.2%(265/270), respectively] and there was no statistical difference. In culture-negative patients with pulmonary tuberculosis, the sensitivity of InnowaveDX was 38.8% (198/511), which was higher than that of Xpert (29.4%, 150/511), and the difference was statistically significant (P<0.001). Taking phenotypic drug-susceptibility testing (DST) as reference, the sensitivity of InnowaveDX to rifampicin resistance was 99.0% (95%CI: 94.7%-100.0%) and the specificity was 94.0%(95%CI: 88.5%-97.4%). With Xpert as the reference, the sensitivity and specificity of InnowaveDX were 97.1% (95%CI: 93.4%-99.1%) and 99.7% (95%CI: 98.4%-100.0%), respectively, and the kappa value was 0.97 (P<0.001). Conclusions: InnowaveDX show a high sensitivity for detecting Mycobacterium tuberculosis, especially in pulmonary tuberculosis patients with a clinical diagnosis and negative culture results. It also showed high sensitivity in detecting rifampicin resistance with DST and Xpert as reference respectively. InnowaveDX is an early and accurate diagnostic tool for TB and drug-resistant TB, particularly suitable for application in low- and middle-income countries.

Characteristics, risk factors and outcomes of critically ill patients with active TB.

BACKGROUND: TB continues to impose a significant healthcare burden despite advancement in diagnostics and increased availability of effective antimicrobials. Recent years have seen a resurgence of the disease in association with increasing life expectancy and use of immunosuppressive therapy. Mortality remains high in TB patients requiring admission to critical care units.METHODS: We conducted a retrospective study in two public hospitals to determine factors associated with mortality in patients with TB requiring critical care admission. All patients aged ≥21 years with a diagnosis of active TB involving any organ system at the time of a critical care admission were eligible. The primary outcome measure was 30-day mortality.RESULTS: Over the study period of 4 years, 148 patients were identified. Overall 30-day mortality was 36.5%. Based on multivariate analysis, factors which independently correlated with 30-day mortality include higher APACHE II (Acute Physiology and Chronic Health Evaluation II) score, acid-fast bacilli smear positivity, initiation of anti-TB treatment prior to critical care admission and need for renal replacement therapy.CONCLUSION: TB in critically ill patients continues to be associated with significant mortality. The factors identified to be associated with poor survival outcomes in our study were largely related to greater disease burden and potential for suboptimal treatment.

[Prevalence and transmission of pyrazinamide-resistant Mycobacterium tuberculosis in Hunan Province,China].

Objective: To provide a scientific reference for the prevention and treatment of pyrazinamide-resistant tuberculosis (PZA-R TB), we analyzed the prevalence and risk factors of pyrazinamide-resistant tuberculosis in Hunan province and described the genotyping and clustering characteristics of the pyrazinamide-resistant Mycobacterium tuberculosis (PZA-R MTB) isolates. Methods: The drug susceptibility test results of first-line anti-tuberculosis drugs including isoniazid (INH), rifampicin (RFP), streptomycin (SM), ethambutol (EMB) and pyrazinamide (PZA), and the characteristics of patients were collected from 3 862 tuberculosis patients in Hunan Chest Hospital (Institute of Tuberculosis Control and Prevention) from January 2016 to December 2018. The prevalence of PZA-R TB was calculated and risk factors were analyzed by univariate and multivariable logistic regression analysis. Two hundred and twelve Mycobacterium tuberculosis isolates selected from June 2017 to June 2018 were genotyped using the 24-loci MIRU-VNTR system. The genetic difference value (h), and the Hunter-Gaston index (HGI) were used to evaluate the resolution and variation for the 24 loci. MIRU-VNTR results were analyzed using BioNumerics 5.0 software to conduct cluster analysis. Clustered isolates were further analyzed by pncA gene sequencing. Results: The rate of PZA-R TB among tuberculosis patients and MDR patients was 14.7%(566/3 862) and 60.5%(511/844), respectively. Multivariable logistic regression analysis showed that patients who were INH mono-resistance and MDR had a higher risk of developing PZA resistance, compared with TB patients who were pan-sensitive to anti-TB drugs (INH, RFP, SM, and EMB). The adjusted OR value (95%CI) was 13.08(5.67-30.18), 298.41(164.88-540.08), respectively, and P values were all less than 0.01. Clustering analysis showed that 65 strains formed 19 clusters, the clustering rate was 30.7%(65/212). Of 19 clusters, eight clusters had at least two isolates with identical pncA mutation types within each cluster. In eight clusters, cluster 4, 6, 16 had four, three, and two patients who lived in the same county, respectively, thus providing probable epidemiological links for the recent transmission of PZA-R Mycobacterium tuberculosis. At least 47.6%(101/212) of PZA drug-resistant TB patients were suggestive of primary drug resistance caused by transmission. Conclusions: The prevalence of PZA-R TB was severe in Hunan province. PZA susceptibility testing should be performed for isolates resistant to any first-line anti-tuberculosis drugs, especially for MDR-MTB isolates. Nearly half of tuberculosis patients were suggestive of primary drug resistance caused by transmission. The prevention and treatment strategy of PZA-R TB should focus on the standardized treatment and management of patients as well as control of the source of infection.

Risk factors associated with catheter-related venous thrombosis: a meta-analysis.

BACKGROUND: So far, the risk factors of catheter-related venous thrombosis (CRVT) are not fully understood. We use evidence-based medicine to find the risk factors of CRVT by pooling the current studies that reported the risk factors of CRVT, aiming to provide guidance for clinical diagnosis and treatment. METHODS: We searched PubMed, Embase, and Cochrane Library from the establishment of the database to July 2021. We included studies that reported the risk factors of CRVT, and we excluded duplicate publications, research without full text, incomplete information or inability to conduct data extraction, animal experiments, reviews, and systematic reviews. STATA 15.1 was used to analyze the data. RESULTS: The pooled results show that history of venous thrombosis (odds ratio [OR] = 3.75, 95% confidence interval [CI]: 1.02-13.85; P = 0.047), cancer (OR = 1.74, 95% CI: 1.17-2.57; P = 0.006), infection (OR = 2.13, 95% CI:1.33-3.42; P = 0.002), and multilumina (OR = 3.34, 95% CI:1.48-7.54; P = 0.004) will significantly increase the occurrence of CRVT. However, there is no significant correlation between sex, congenital heart disease, bedridden state, sepsis, mechanical ventilation, anticoagulation therapy, insertion site (left), and CRVT. CONCLUSION: Our research results indicate that history of venous thrombosis, cancer, infection and multilumina are possible risk factors for CRVT, and corresponding preventive measures should be taken clinically.

[Restratifying the prognosis of acute myeloid leukemia patients with CEBPA double mutations based on CSF3R mutations and measurable residual disease].

Objective: This investigation aims to assess the impact of CSF3R mutations and the presence of measurable residual disease (MRD) on the prognosis of patients with CEBPA double mutations who have acute myeloid leukemia (AML) . Methods: The prognostic significance of these two factors was examined in the present study, which included 66 patients with complete genetic mutations and sequential MRD information. Results: Following the second course of chemotherapy, the MRD status and CSF3R mutations of these patients were linked to their long-term prognosis. CSF3R mutated patients showed inferior relapse-free survival (RFS) (5-year RFS: 15.2% vs 38.7% , P=0.006) and overall survival (OS) (5-year OS: 18.2% vs 60.6% , P=0.038) compared with those with wild-type CSF3R. After the second course of chemotherapy, patients with negative MRD had an RFS of 64 months and an OS of not reaching, which was significantly longer than that of patients with positive MRD (15 and 48 months, and the P value were 0.004 and 0.050, respectively) . CSF3R mutations (HR=0.317, 95% CI 0.129-0.779, P=0.012) , WT1 mutations (HR=0.304, 95% CI 0.115-0.804, P=0.016) , and NRAS mutations (HR=0.153, 95% CI 0.061-0.385, P<0.001) were all independently associated with a poor prognosis for RFS, and CSF3R mutations and positive MRD tended to be independently associated with a poor prognosis for OS, according to the results of a Cox proportional-hazards model analysis (P values were 0.071 and 0.088, respectively) . The patients were divided into three groups based on their CSF3R mutation status and MRD status following treatment: wide-type CSF3R and negative MRD, mutated CSF3R or positive MRD, and mutated CSF3R and positive MRD, which showed significantly different RFS (P<0.001) and OS (P=0.006) . Conclusion: Both CSF3R mutations and positive MRD were associated with poor outcome in AML patients with CEBPA double mutations. An integrity model based on these two factors may be beneficial for accurately evaluating the prognosis of these patients.

Hyperglyceridaemia-induced acute pancreatitis in pregnancy: Experience from a tertiary hospital in Singapore.

Acute pancreatitis (AP) is a serious condition that can occur suddenly in pregnancy. We present a case of sudden onset of epigastric pain with severely deranged serum triglyceride levels in a 32-year-old Vietnamese primigravida with no significant past medical history in the Singapore General Hospital. The patient was managed in the intensive care unit, with plasmapheres and intravenous insulin and was eventually a healthy term foetus was delievered via ceasarian section. This case showcased multidisciplinary co operation between the obstetrics, anaesthetic, endocrinology and intensive care team and serves as a reminder to consider this rare condition for future similar presentations.

[The derivation and vulidafion of a prediction rule for hypoxemia in infants with Pierre Robin sequence after weaning].

Objective: To set up a prediction scoring system for the hypoxemia in infants with Pierre Robin sequence after weaning and evaluate its clinical value. Methods: Data of consecutive patients from November 2016 to June 2019, who underwent mandibular distraction osteogenesis in Guangzhou Women and Children's Medical Center, were retrospectively analyzed (n=148). All the cases were divided into two groups according to the appearance of hypoxemia after weaning. They were randomly divided into the derivation cohorc (2/3,n=100) and the validation cohort (1/3,n=48). Single factor and multiple logistic regression analysis were used to select the independent risk factors related to hypoxemia and establish a prediction model. A prediction scoring system was developed in accordance with assigning of the value of each variable ß in the model. Internal verification of scoring system by validation population. Data of consecutive patients from July 2019 to November 2019, who underwent mandibular distraction osteogenesis, were prospectively analyzed (n=26). The diagnostic accuracy were conducted to evaluate the clinical value of the scoring system. Results: The logistic regression demonstrated that age at operation, pulmonary infection and the length of distraction less than 5 mm at weaning were the independent risk factors for hypoxemia. The P value of logistic regression model in Hosmer and Lemeshow goodness of fit test was 0.848, and a prediction scoring system was established accordingly. The area under the ROC curve of the scoring system was 0.890, and the optimum critical value was 53. The sensitivity, specificity, accuracy of the model were 78.6%(11/14),86.1%(74/86), 85.0%(85/100) respectively. The predictive effectiveness of the scoring system in the retrospective validation population was similar to that in the modeling population. 26 patients were included in the prospective analysis. The area under ROC curve of the scoring system was 0.870. The sensitivity, specificity and accuracy were 80.0%(5/6),95.0%(20/21), 96.1%(25/26) respectively. Conclusion: The prediction scoring system established in the study are efficacious for the hypoxemia in infants with Pierre Robin sequence after weaning.

Expression of cytosolic and thiolated proteome of Musca domestica larvae under oxidative challenge.

The study was aimed to investigate the expression of cytosolic and thiolated proteins of Musca domestica larvae under oxidative stress. Proteins from acute treatment of hydrogen peroxide (LC50 = 21.52% (v/v)) on 3rd stage larvae of housefly were extracted and purified using an activated Thiol Sepharose® for thiolated protein purification. Two dimensional gel electrophoresis was used for visualizing and analyzing expression of cytosolic and thiolated proteins. Protein spots with more than 5 fold of expression change were identified using liquid chromatography- tandem mass spectrometry (LC-MS/MS). The cytosolic proteins were actin, tropomyosin, ubiquitin, arginine kinase, pheromone binding protein/general odorant binding protein, and ATP: guanidino phosphotransferase. The thiolated proteins with more than 5 fold change in expression as an effect to the acute treatment were fructose bisphosphate aldolase, short chain dehydrogenase and lactate/malate dehydrogenase. The proteins identified in the study should provide vital information for future reference in oxidative stress defence and response occurring in houseflies.

Expression of cytosolic and thiolated proteome of Musca domestica larvae under oxidative challenge

@#The study was aimed to investigate the expression of cytosolic and thiolated proteins of Musca domestica larvae under oxidative stress. Proteins from acute treatment of hydrogen peroxide (LC50 = 21.52% (v/v)) on 3rd stage larvae of housefly were extracted and purified using an activated Thiol Sepharose® for thiolated protein purification. Two dimensional gel electrophoresis was used for visualizing and analyzing expression of cytosolic and thiolated proteins. Protein spots with more than 5 fold of expression change were identified using liquid chromatography- tandem mass spectrometry (LC-MS/MS). The cytosolic proteins were actin, tropomyosin, ubiquitin, arginine kinase, pheromone binding protein/general odorant binding protein, and ATP: guanidino phosphotransferase. The thiolated proteins with more than 5 fold change in expression as an effect to the acute treatment were fructose bisphosphate aldolase, short chain dehydrogenase and lactate/malate dehydrogenase. The proteins identified in the study should provide vital information for future reference in oxidative stress defence and response occurring in houseflies.

[Impact of KIT D816 mutation on salvage therapy in relapsed acute myeloid leukemia with t(8;21) translocation].

Objective: To evaluate the impact of KIT D816 mutation on the salvage therapy in relapsed acute myeloid leukemia (AML) with t(8;21) translocation. Method: The characteristics of the first relapsed AML with t(8;21) translocation from 10 hospitals were retrospectively collected, complete remission (CR(2)) rate after one course salvage chemotherapy and the relationship between KIT mutation and CR(2) rate was analyzed. Results: 68 cases were enrolled in this study, and 30 cases (44.1%) achieved CR(2). All patients received KIT mutation detection, and KIT D816 mutation was identified in 26 cases. The KIT D816 positive group had significantly lower CR(2) compared with non-KIT D816 group (23.1% vs 57.1%, χ(2)=7.559, P=0.006), and patients with longer CR(1) duration achieved significantly higher CR(2) than those with CR(1) duration less than 12 months (74.1% vs 31.9%, χ(2)=9.192, P=0.002). KIT D816 mutation was tightly related to shorter CR(1) duration. No significant difference of 2 years post relapse survival was observed between KIT D816 mutation and non-KIT D816 mutation group. Conclusion: KIT D816 mutation at diagnosis was an adverse factor on the salvage therapy in relapsed AML with t(8;21) translocation, significantly related to shorter CR1 duration, and can be used for prediction of salvage therapy response. KIT D816 mutation could guide the decision-making of salvage therapy in relapsed AML with t(8;21) translocation.

[Effect of 1q21 amplification on bortezomib therapeutic response and prognosis of newly diagnosed multiple myeloma patients].

Objective: To investigate the effect of 1q21 amplification (1q) on the therapeutic response and prognosis of bortezomib(Btz) in the treatment of newly diagnosed multiple myeloma (MM) patients. Methods: A total of 180 newly diagnosed MM were included for analyses of clinical characteristics, cytogenetics, objective response rate (ORR), progression-free survival (PFS) and overall survival (OS), retrospectively. Gene expression profiling (GEP) was analyzed using publicly available R2 platform. Results: ① In 180 patients, 1q was found in 51.1% cases. Of them, 174 patients had complete follow-up data, including 88 cases with 1q and 86 without 1q (non-1q). ②Incidence of 1q was positively associated with percentage of IGH rearrangement (72.2%, P=0.017) and 1p deletion (1p) (27.8%, P=0.040). ③ The median PFS was 15.0 and 20.3 months for the 1q group and non-1q group, and the median OS was 29.4 and 44.0 months, respectively. Both PFS and OS of 1q group was significantly shorter than those of the non-1q group (P=0.029 and 0.038, respectively). Multivariate analysis further revealed that 1q was an independent prognostic factor for both PFS (HR=1.910, 95% CI 1.105-3.303, P=0.020) and OS (HR=2.353, 95% CI 1.090-5.078, P=0.029). ④ In 91 evaluable cases with 1q, very good partial remission (VGPR) rate was higher after treatment with Btz than those without Btz (62.1% vs 40.0%, P=0.032). Of note, the patients with 1q who received auto-HSCT after induction with Btz had significantly longer PFS than those without auto-HSCT (19 months vs 13 months, P=0.048). ⑤GEP analysis revealed that 1q21 amplification predominantly up-regulated expression of >50% genes within 1q21 region, and also altered expression of 28% genes in chromosome 1 and 10% genes in whole genome, particularly related to DNA repair and cell cycle. Conclusions: 1q is an independent adverse prognostic factor in patients with newly diagnosed MM. It is often associated with 1p deletion and IGH rearrangement. Patients with 1q respond well to Btz-based regimen, but they fail to gain long-term benefit from this treatment itself. However, auto-HSCT following Btz induction might improve survival of patients with 1q, suggesting a potential strategy to treat this high-risk subset of MM. GEP analysis warrants further attention in understanding the mechanisms underlying the high-risk of 1q.

[Effect of TSC2 gene expression downregulation by lentivirus induced RNA interference on U937 cell line and its mechanism].

Objective: To investigate the effect of biology and mTOR pathway activity of down-regulated TSC2 gene expression on U937 leukemia cells. Methods: Gene expression was down-regulated by lentivirus induced RNA interference on TSC2 high expressed U937 cell line; the proliferation, apoptosis and differentiation were detected by CCK-8 assay, colony formation assay and flow cytometry; the gene expression level and protein kinase activity were detected by qRT-PCR and Western blot. Results: Down-regulated expression of TSC2 gene promoted U937 cell proliferation and colony formation ability (P<0.05) . The proportion in G(0)/G(1) phase of TSC2 down-regulated U937 cell was much lower than that of the control cells [ (52.53±3.75) % vs (75.10±4.33) %, t=6.829, P=0.002], the S phase [ (22.43±1.00) % vs (15.47±1.20) %, t=-5.581, P=0.019] and G(2)/M phase [ (25.03±4.34) % vs (14.33±0.91) %, t=-5.413, P=0.013] was remarkably higher than that of the control cells (P<0.05) . There were no statistically significant differences in cell apoptosis and differentiation (P>0.05) . Down-regulation of TSC2 led to the increased activity of mTOR, 4EBP1 and S6K1, but did not influence the activity of AKT. The expressions of proliferation related cyclinD1, c-myc and PTEN were also up-regulated after TSC2 silenced, but the expressions of P27KIP and BCL-XL were not changed. Conclusion: Downregulation of TSC2 could promote the proliferation of U937 cells through up-regulation of mTOR activity.

Body height and the spread of spinal anaesthesia for caesarean section: a prospective controlled trial.

BACKGROUND: No conclusive evidence exists on the effect of patient height on the spread of spinal anaesthesia. Our aim was to measure the ED50 and ED95 values of intrathecal ropivacaine in taller and shorter patients, and thus investigate the hypothesis that the spinal dose requirement in shorter patients is lower than that in taller patients undergoing caesarean section. METHODS: In this study, 270 pregnant women were assigned to the taller (Group T) or shorter group (Group S) based on their heights. Subjects in both groups were further randomly assigned to one of nine subgroups based on the dosage of intrathecal isobaric ropivacaine to be administered (7, 8, 9, 10, 11, 12, 13, 14 or 15 mg respectively). RESULTS: The ED50 and ED95 values of ropivacaine were 9.24 mg and 13.36 mg in Group S, and 10.11 mg and 14.63 mg in Group T, with no inter-group difference (P = 0.886). There was a significant inter-group difference in the incidence of hypotension and the changes in mean arterial pressure after spinal anaesthesia using 15 mg ropivacaine. The dose of ephedrine administered in Group S was higher than that in Group T when 15 mg ropivacaine was administered (P = 0.031). CONCLUSION: The taller and shorter patients did not respond differently to modest intrathecal doses of ropivacaine. However, a larger dose of ropivacaine was associated with an increased incidence of hypotension in shorter patients compared to that in taller patients.